BstNI NEB#R0168

Protocol tips

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	Digestion of 157 bp PCR product of RAD51 (rs1801320) was dispatched with BstNI (NEB,USA) by incubating at 60° C for 4 hours which produced two fragments 86 bp and 71 bp in case of G/G allele while for C/C allele it produced only one fragment of 157 bp.

Protocol tips
Digestion of 157 bp PCR product of RAD51 (rs1801320) was dispatched with BstNI (NEB,USA) by incubating at 60° C for 4 hours which produced two fragments 86 bp and 71 bp in case of G/G allele while for C/C allele it produced only one fragment of 157 bp.

Publication protocol

"Genomic DNA was separated from the blood specimens assembled from all patients by the help of DNA extraction mini kit, Favorgen, Taiwan. Genomic
DNA amplification was executed using the predesigned
forward and reverse primers. PCR was done by following
the standard protocol mentioned in published method
(Shabnaz et al., 2015). In case of both RAD51 (rs1801320)
and XRCC2 (rs3218536), PCR-Restriction Fragment
Length Polymorphism (RFLP) approach was applied
to genotype. PCR product of 157 bp and 234 bp were
obtained for RAD51 (rs1801320) and XRCC2 (rs3218536)
SNPs respectively. PCR amplifications conditions are
given in Table 3. Digestion of 157 bp PCR product of
RAD51 (rs1801320) was dispatched with BstNI (NEB,
USA) by incubating at 60° C for 4 hours which produced
two fragments 86 bp and 71 bp in case of G/G allele while
for C/C allele it produced only one fragment of 157 bp.
Restriction enzyme Hph1 (NEB, USA) (37° C overnight)
bisected the T/T allele of XRCC2 (rs3218536) giving two
fragments of 147 bp and 87 bp but didn’t divide the C/C
allele that is only one fragment 234 bp was observed.
These DNA fragments were blemished with ethidium
bromide and pictured on 2% agarose gel electrophoresis"

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